Abstract
An anti-hepatitis B surface antigen (HBsAg), single-chain Fv antibody fragment (scFv) with a 6-histidine N-terminal tag was produced in cultured transgenic tobacco cells. Western blot and antigen-specific chromatography showed high levels of biologically active scFv in the culture supernatant (1 mg l−1) and in cells (5 mg kg−1). A simple one-step scFv purification was developed using immobilized metal ion affinity chromatography.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ayala M, Fernández-de-Cossío ME, Canaán-Haden L, Balint RF, Larrick JW, Gavilondo JV (1995) Variable region sequence modulates periplasmic export of a single chain Fv antibody fragment in E. coli. BioTechniques 18: 832-842.
Canaán-Haden L, Ayala M, Fernández-de-Cossio ME, Pedroso I, Rodés L, Gavilondo JV (1995) Purification and application of a single chain Fv antibody fragment specific to hepatitis B virus surface antigen. BioTechniques 19: 606-614.
Firek S, Draper J, Owen MRL, Gandacha A, Cockburn B, Whitelam G (1993) Secretory of a functional single-chain Fv protein in transgenic tobacco plants and cell suspension culture. Plant Mol. Biol. 23: 861-870.
Fischer R, Liao YC, Drossard J (1999) Affinity-purification of a TMV-specific recombinant full-size antibody from a transgenic tobacco suspension culture. J. Immunol. Meth. 226: 1-10.
Hein M, Tang Y, McLeod DA, Janda KD, Hiatt A (1991) Evaluation of immunoglobulins from plant cells. Biotechnol Prog. 7: 455-461.
Larrick JW, Yu L, Chen J, Jaiswal S, Wycoff K (1998) Production of antibodies in transgenic plants. Res. Immunol. 149: 603-608.
López A, ZaldÚa Z, Pimentel E, García M, García R, Mena J, Morán R, Selman G (1996) Modification of sporamin gene from sweet potato with a synthetic DNA fragment. Nucleotide sequence and expression in Escherichia coli. Biotecnología Aplicada 13: 265-270.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15: 473-497.
Porath J (1992) Immobilized metal ion affinity chromatography. Prot. Express. Purif. 3: 263-281.
van Engelen FA, Schouten A, Molthoff JW, Roosien J, Salinas J, Dirkse WG, Schots A, Bakker J, Gommers FJ, Jongsma MA, Bosch D, Stiekema WJ (1994) Coordinate expression of antibody subunit genes yields high levels of functional antibodies in root of transgenic tobacco. Plant Mol. Biol. 26: 1701-1710.
Zambrysky P, Joos H, Gentello J, Leemans J, Van Montagu M, Schell J (1983) Ti-plasmid vector for introduction of DNA into plant cells without altering their normal regeneration capacity. EMBO J. 2: 2143-2150.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ramírez, N., Lorenzo, D., Palenzuela, D. et al. Single-chain antibody fragments specific to the hepatitis B surface antigen, produced in recombinant tobacco cell cultures. Biotechnology Letters 22, 1233–1236 (2000). https://doi.org/10.1023/A:1005605214703
Issue Date:
DOI: https://doi.org/10.1023/A:1005605214703